Cryopreservation of zygotic embryos of Podophyllum hexandrum Royle, an endangered medicinal plant, by vitrification and v cryo-plate techniques.

BACKGROUND: Podophyllum hexandrum is a highly endangered valuable medicinal plant of the Himalayas belonging to family Berberidaceae. This plant needs conservation efforts due to the over-exploitation and unscrupulous harvesting from the wild because of its ever-increasing demand. OBJECTIVE: To establish a long-term cryopreservation method for Podophyllum hexandrum using two techniques: Vitrification and V Cryo-plate. MATERIALS AND METHODS: Zygotic embryos were cryopreserved using vitrification and V cryo-plate by optimization of parameters including preculture time, loading time and PVS2 dehydration time. Recovery of zygotic embryos was performed on different regrowth media for plantlet formation. RESULTS: With V cryo-plate, 90% regrowth was obtained as compared to 73.3% with vitrification. V Cryo-plate conditions were pre-culture of zygotic embryos in 0.3 M sucrose for 4 days, treatment in loading solution with 0.8 M sucrose for 20 min, dehydration in PVS2 for 50 min, LN exposure, unloading in 1.2 M sucrose for 20 min and transfer of zygotic embryos to regrowth medium for recovery. During recovery, the maximum number of shoots (4.2) and highest shoot length (5.1 cm) were observed on regrowth medium with 1.5 mg per liter BAP and 0.1 mg per liter IAA (R7). CONCLUSION: Zygotic embryos of Podophyllum hexandrum were cryopreserved with 90% regrowth using a V cryoplate technique and plantlets were produced directly after cryopreservation. Doi: 10.54680/fr23410110712.

Cytotoxic Potential and Molecular Characterization of Fungal Endophytes from Selected High Value Medicinal Plants of the Kashmir Valley - India.

The present study explores the fungal endophytes from selected high value medicinal plants to check their activities at in-vitro and in-vivo level. The in-vitro cytotoxicity of selected endophytes revealed potent growth inhibition against human cancer cell lines of leukemia (THP-1), lung (A549), prostate (PC-3), colon (Caco-2), neuroblastoma (IMR-32) and breast (MCF-7) at a concentration of 100 µg/ml. Among them the endophytic strains I. e., IIIM2, IIIM3, IIIM7 and IIIM8 showed most significant growth inhibition against colon (Caco-2), prostate (PC-3), lung (A549) and leukemia (THP-1) cancer cell lines. At the in-vivo level maximum (58.95%) tumor growth inhibition was documented with the extract of IIIM2 against Ehrlich Ascites Carcinoma mouse modal. All the potent fungal endophytic strains were characterized using ITS 4 and ITS 5 region sequencing and phylogenetic analysis was ascertained among them. This paper confirms the 2 elite endophytic fungal strains, IIIM2 and IIIM8, have the potential to act as a source of new anticancer compounds.

Cloning, sequencing and partial characterisation of sorbitol transporter (srlT) gene encoding phosphotransferase system, glucitol/sorbitol-specific IIBC components of Erwinia herbicola ATCC 21998.

A DNA fragment of approximately 1500 bp, harbouring the sorbitol transport gene (srlT), was amplified from the chromosomal DNA of Erwinia herbicola ATCC 21998 by PCR and cloned in Escherichia coli JM109. Degenerate oligonucleotide primers used were designed based on the conserved regions in the gene sequences within the gut operon of E. coli (Gene Bank accession no. J02708) and the srl operon of Erwinia amylovora (Gene Bank accession no. Y14603). The cloned DNA fragment was sequenced and found to contain an open reading frame of 1473 nucleotides coding for a protein of 491 amino acids, corresponding to a mass of 52410 Da. The nucleotide sequence of this ORF was highly homologous to that of the gutA gene of Escherichia coli gut operon, the srlE gene of Shigella flexrni and the sorbitol transporter gene sequence of Escherichia coli K12 (Gene Bank accession nos. J02708, AE016987 and D90892 respectively). The protein sequence showed significant homology to that of the phosphotransferase system i.e. the glucitol/sorbitol-specific IIBC components of Escherichia coli and Erwinia amylovora (P56580, O32522). The cloned DNA fragment was introduced into a pRA90 vector and the recombinant was used for developing srlT mutants of Erwinia herbicola, by homologous recombination. Mutants obtained were unable to grow on minimal medium with sorbitol. The insertion of the pRA90 vector inside the srlT gene sequence of the mutants was confirmed by DNA-DNA hybridisation.